斑点免疫金渗滤法检测抗荚膜组织胞浆菌抗体的研究

目的 研究快速检测荚膜组织胞浆菌抗体的方法.方法 荚膜组织胞浆菌素纯蛋白衍生物(P-HTPM)为包被抗原,以葡萄球菌A蛋白(SPA)标记的免疫胶体金为检测标记物,采用斑点免疫金渗滤试验(DIGFA)检测荚膜组织胞浆菌(HC)免疫小鼠血清中抗HC抗体以及其他真菌免疫血清中抗体.结果 DIGFA检测20份HC小鼠免疫血清,结果全为阳性,与ELISA检测结果一致,敏感性为100%.检测8份正常小鼠血清出现1份假阳性,特异性为87.5%.检测其他真菌免疫血清结果为:念珠菌属和马尔尼菲青霉菌免疫血清结果全为阴性;8份皮炎芽生菌血清出现1份阳性,9份副球孢子菌血清出现2份阳性.结论 斑点免疫金渗滤法检测抗HC抗体有较高的敏感性和特异性,快速简便.经临床标本扩大验证后将对荚膜组织胞浆菌病的快速诊断有应用价值.

Detection of anti-histoplasma antibody in sera of immunized mice by dot immunogold filtration assay

Objective To study a new and rapid method for detection of anti-histoplasma antibody by dot immunogold filtration assay (DIGFA). Methods DIGFA was developed by coating purified protein derivative of histoplasmin (P-HTPM) on nitrocellulose membrane as membrane antigen and labeling SPA with colloidal gold. Anti-histoplasma antibodies in sera from mice immunized with Histoplasma were detected by DIGFA. Results All immunizedsera with Histoplasma were positive by DIGFA as well as the results detected by ELISA. The sera immunized respectively with Monilia and Penicillium marneffei were negtive by DIGFA while 1 of 8 immunizedsera with Blastomyces dermatitidis and 2 of 9 immunizedsera with Paracoccidioides brasiliensis were found to be positive. Conclusion DIGFA was a valuable and rapid method to detect histoplasmaantibody in serum.